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Antibody and Protein Labeling


Immunostain
Labeling an antibody with a specific tag has become an important and routine procedure in biological sciences and medical research. Tags, which range from visual markers such as fluorescent dyes to hapten molecules like biotin, can be used to enhance the visibility of immune complexes and protein-protein interactions or can be used in the isolation and purification of a protein.

When determining which type of label to choose, focus on the downstream application for which the antibody will be used. Fluorescent tags are used predominantly in cellular imaging, flow cytometry, fluorescent blotting, and immunofluorescence applications. In contrast, enzymatic tags, such as HRP and AP, are commonly used for blotting, ELISAs, and immunohistochemistry applications. Hapten tags such as biotin are the more versatile of the possible tag options as they can be used in protein isolation, purification, and any of the aforementioned applications.

AAT Bioquest offers a comprehensive suite of antibody and protein labeling kits to choose from. Designed for the novice scientist in mind, these kits have the easiest protocols and require the least hands-on experience.

 



Antibody Labeling Kits At-A-Glance



Antibody Labeling Kit ReadiLink™ Kits ReadiLink™ xtra Kits Buccutite™ Kits
Amount of antibody labeled per reaction 50 µg 50 µg 25 µg, 50 µg or 1 mg
Labeling Chemistry Targets free amine groups using amine-reactive dyes Targets free amine groups using amine-reactive dyes Uses two proprietary linkers to facilitate protein-protein conjugation
Available labels iFluor™ dyes, mFluor™ dyes, Alexa Fluor® equivalent AF dyes, FITC, Cy3, Cy5, Cy7, Biotin iFluor™ dyes, Alexa Fluor® equivalent AF dyes, FITC, Cy3, Cy5, Cy7 PE, APC, Tandem Dyes, HRP, AP
Type of bond between label and antibody Covalent Covalent Covalent
Compatible with BSA or other stabilizers? No Yes No
Requires purification? No No No
Time required for conjugation 1 hr 1 hr 2 hrs
Hands-on-time ~10 minutes ~10 minutes ~15 minutes
Batch-to-batch variation Minimal Minimal Minimal
Applications¹ Imaging, FC, IF, WB Imaging, FC, IF, WB FC, IF, IHC, WB, ELISA
  1. FC = flow cytometry; IF = immunofluorescence; IHC = immunohistochemistry; WB = Western blot; ELISA = enzyme-linked immunosorbent assay.

 


AAT Bioquest ReadiLink™ Rapid Antibody Labeling Kits provide a quick and convenient method to label microscale volumes of antibodies with our superior iFluor™ dyes, mFluor™ dyes, or other labels. The unique chemistry of ReadiLink™ kits enables researchers to effortlessly label and recover 100% of their antibodies without a purification step. Since ReadiLink™ conjugates are covalently labeled, they are stable for long-term storage. They are ideal for demanding applications, including fluorescent microscopy, multi-color imaging, multiplex flow cytometry, primary detection, IHC, FISH, ELISAs, and more.

Key features of the ReadiLink™ Rapid Antibody Labeling Kits

  • Label monoclonal antibodies, polyclonal antibodies, or other proteins
  • Quick and simple two-step labeling protocol, no chemistry or conjugation experience necessary
  • Useful for labeling 100 µg of antibody (two separate labeling reactions of 50 µg antibody)
  • 100% antibody recovery, NO purification of the product is required
  • Conjugates ready to use in 1 hr, with as little as ~10 minutes of hands-on time
  • Choice of iFluor™ dyes, mFluor™ dyes, Alexa Fluor® equivalent AF dyes, FITC, Cy3, Cy5, Cy7, BSA, KLH or biotin
  • Avoid using any amine-containing buffers (e.g., Tris or glycine)
  • Stabilizing proteins (e.g., BSA or gelatin) must be removed before labeling; for antibody labeling kits compatible with BSA, use ReadiLink™ xtra Rapid Antibody Labeling Kits

Convenient Design for Maximum Results



ReadiLink™ antibody labeling workflow (figure drawn in BioRender).
ReadiLink™ Rapid Antibody Labeling Kits utilize amine-reactive labels to prepare covalently labeled conjugates. These reactive labels selectively target and bind to primary amines, such as lysine residues, on the antibody of interest. To label your antibody, mix your antibody solution (concentration ~1 mg/mL) with the ReadiLink™ reaction buffer, add the mixture to the vial of pre-formulated label, and incubate for 30 to 60 minutes. Lastly, add the TQ™-Dyed Quench Buffer to your mixture and mix well. This part is key as it eliminates both background interference and the column purification step. After 10 minutes of incubation, your conjugates are ready to use.

High-performance alternative to Lightning-Link® Antibody Labeling Kits



Flow cytometry analysis of human PBMC cells. Anti-human CD4 antibody prepared with ReadiLink™ Rapid iFluor488 Antibody Labeling Kit (Cat No. 1255) or Lightning-Link® Rapid Alexa Fluor ® 488. The fluorescence signal was monitored using the ACEA NovoCyte flow cytometer in the FITC channel.
Labeling Kit CD4(SK3)- Conjugate Stain Index
ReadiLink™ Kit CD4-iFluor™ 488 Conjugate 43
Lightning-Link® Kit CD4-Alexa Fluor® 488 Conjugate 26

 

Table 1. ReadiLink™ Rapid Antibody Labeling Kit Selection Guide

Fluorophore
Fluorophore Equivalent
Ex/Em (nm)
Imaging
Flow Cytometry
Cat No.
mFluor™ Violet 420Cascade® Blue398/411 -Yes1105
iFluor™ 350AMCA, Alexa Fluor® 350345/442Yes -1220
mFluor™ Violet 450Pacific Blue®403/454 -Yes1100
mFluor™ Violet 510414/508 -Yes1110
iFluor™ 488FITC, Alexa Fluor® 488491/514YesYes1255
FITC494/520YesYes1299
mFluor™ Violet 540Pacific Orange®405/537 -Yes1114
Cy3555/565Yes -1290
iFluor™ 555Cy3, Alexa Fluor® 555552/567Yes -1227
mFluor™ Blue 570R-PE553/570 -Yes1120

 


ReadiLink™ xtra Rapid Antibody Labeling Kits with BSA compatibility takes labeling primary antibodies to a whole new level of convenience. Utilizing an optimized labeling protocol combined with vials of specially formulated labels, ReadiLink™ xtra kits enable rapid and reproducible labeling of microgram quantities of a primary antibody in the presence of common buffer additives and stabilizing proteins. In just two simple steps and without column purification, covalently label antibodies suitable for use in any immuno-based application.

Key features of the ReadiLink™ xtra Rapid Antibody Labeling Kits

  • Compatible with BSA and other stabilizing proteins
  • Label monoclonal antibodies, polyclonal antibodies, or other proteins
  • Labels 100 µg of antibody (two separate labeling reactions of 50 µg antibody)
  • 100% antibody recovery, NO purification of the product is required
  • Conjugates ready to use in 1 hr, with as little as ~10 minutes of hands-on time
  • Choice of iFluor™ dyes, mFluor™ dyes, Alexa Fluor® equivalent AF dyes, FITC, Cy3, Cy5 or Cy7

 

Table 2. ReadiLink™ xtra Rapid Antibody Labeling Kits

Name
Ex (nm)
Em (nm)
Unit Size
Cat No.
ReadiLink™ xtra Rapid iFluor™ 350 Antibody Labeling Kit3444482 Labelings1950
ReadiLink™ xtra Rapid iFluor™ 488 Antibody Labeling Kit4915162 Labelings1955
ReadiLink™ xtra Rapid AF488 Antibody Labeling Kit4995202 Labelings1978
ReadiLink™ xtra Rapid FITC Antibody Labeling Kit4915162 Labelings1970
ReadiLink™ xtra Rapid iFluor™ 555 Antibody Labeling Kit5565692 Labelings1958
ReadiLink™ xtra Rapid AF555 Antibody Labeling Kit5535682 Labelings1980
ReadiLink™ xtra Rapid iFluor™ 594 Antibody Labeling Kit5876032 Labelings1960
ReadiLink™ xtra Rapid AF594 Antibody Labeling Kit5906182 Labelings1982
ReadiLink™ xtra Rapid iFluor™ 647 Antibody Labeling Kit6546692 Labelings1963
ReadiLink™ xtra Rapid AF647 Antibody Labeling Kit6506712 Labelings1985

 

Buccutite™ Protein to Protein Labeling Kits


Buccutite™ crosslinking technology offers a trouble-free way to covalently label proteins and antibodies with horseradish peroxidase (HRP), alkaline phosphatase (AP), or a phycobiliprotein. Compared to the conventional techniques for protein-protein conjugation, such as the SMCC chemical labeling method, Buccutite™ conjugation is significantly more robust and less demanding. These kits allow optimal labeling of microgram amounts of antibodies (as little as 25 µg per labeling reaction) in less than two hours, with no purification necessary and 100% yield. With Buccutite™ labeling kits, routinely prepare covalently-labeled conjugates that will withstand the rigorous washing techniques generally required in a typical immunoassay (e.g., ELISA, IHC, and Western blot) or flow cytometry application.

Key features of the Buccutite™ Rapid Antibody Labeling Kit

  • Label 25 µg up to 1 mg of antibody
  • Choice of labels include HRP, AP, phycoerythrin (PE), allophycocyanin (APC), PerCP, PE-tandem dyes, and APC tandem dyes
  • Less than 2 hours of labeling time, with no purification required
  • 100% antibody recovery

Learn More


High-performance alternative to Lightning-Link® Antibody Labeling Kits



Flow cytometry analysis of human PBMC cells. Anti-human CD4 antibody prepared with Buccutite™ Rapid PE Antibody Labeling Kit (Cat No. 1310) or Lightning-Link® PE. The fluorescence signal was monitored using the ACEA NovoCyte flow cytometer in the FITC channel.
Labeling Kit CD4(SK3)- Conjugate Stain Index
Buccutite™ Kit CD4-PE 488 Conjugate 55
Lightning-Link® Kit CD4-PE Conjugate 28

 

Table 3. Buccutite™ Rapid Antibody Labeling Kits

Name
Ex (nm)
Em (nm)
Unit Size
Cat No.
Buccutite™ Rapid Protein Crosslinking Kit *Microscale Optimized for Crosslinking 100 ug Antibody Per Reaction*--2 Labelings1315
Buccutite™ Peroxidase (HRP) Antibody Conjugation Kit *Optimized for Labeling 1 mg Protein*--1 Kit5504
Buccutite™ Peroxidase (HRP) Antibody Conjugation Kit *Optimized for Labeling 100 ug Protein*--1 Kit5503
Buccutite™ Peroxidase (HRP) Antibody Conjugation Kit *Optimized for Labeling 25 ug Protein*--2 Labelings5505
Buccutite™ Rapid PE Antibody Labeling Kit *Microscale Optimized for Labeling 100 ug Antibody Per Reaction*5655752 Labelings1310
Buccutite™ Rapid PE Antibody Labeling Kit *Microscale Optimized for Labeling 25 ug Antibody Per Reaction*5655752 Labelings1312
Buccutite™ Rapid PE-Texas Red Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 100 ug Antibody Per Reaction*5656002 Labelings1318
Buccutite™ Rapid PE-Texas Red Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 25 ug Antibody Per Reaction*5656002 Labelings1343
Buccutite™ Rapid PE-Cy5 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 100 ug Antibody Per Reaction*5656742 Labelings1322
Buccutite™ Rapid PE-Cy5 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 25 ug Antibody Per Reaction*5656742 Labelings1340



Product Ordering Information



Table 4. Ordering Info for Readilink Antibody Labeling Kits Products

Cat#
Product Name
Unit Size
Price
Ex
Em
1100ReadiLink™ Rapid mFluor™ Violet 450 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction*2 Labelings153403454
1105ReadiLink™ Rapid mFluor™ Violet 420 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction*2 Labelings153398411
1110ReadiLink™ Rapid mFluor™ Violet 510 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction*2 Labelings153414508
1114ReadiLink™ Rapid mFluor™ Violet 540 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction*2 Labelings153405537
1120ReadiLink™ Rapid mFluor™ Blue 570 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction*2 Labelings153553570
1123ReadiLink™ Rapid mFluor™ Green 620 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction*2 Labelings153522617
1130ReadiLink™ Rapid mFluor™ Red 700 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction*2 Labelings153657700
1131ReadiLink™ Rapid mFluor™ Red 780 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction*2 Labelings153629780
1220ReadiLink™ Rapid iFluor™ 350 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction*2 Labelings153345442
1227ReadiLink™ Rapid iFluor™ 555 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction*2 Labelings153552567